Store at
-20ºC
#15071
Bcl-2 (124) Mouse mAb
www.cellsignal.com
Support: 877-678-TECH (8324)
www.cellsignal.com/support
100 µl (10 western blots)
Orders: 877-616-CELL (2355)
[email protected]
rev. 05/22/15
Entrez-Gene ID #596
UniProt ID #P10415
For Research Use Only. Not For Use In Diagnostic Procedures.
Applications
W, IP, IHC-P, F
Endogenous
Species Cross-Reactivity*
Molecular Wt.
Isotype
H
26 kDa
Mouse IgG1**
Storage: Supplied in 10 mM sodium HEPES (pH 7.5), 150
mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02%
sodium azide. Store at –20°C. Do not aliquot the antibody.
*Species cross-reactivity is determined by western blot.
Specificity/Sensitivity: Bcl-2 (124) Mouse mAb recognizes
endogenous levels of total Bcl-2 protein.
Source/Purification: Monoclonal antibody is produced by
immunizing animals with a synthetic peptide corresponding to
residues surrounding Gly47 of human Bcl-2 protein.
Background References:
(1) M
urphy, K.M. et al. (2000) Cell Death Differ 7, 102-11.
(2) Z hu, L. et al. (1999) J Biol Chem 274, 33267-73.
(3) M
aundrell, K. et al. (1997) J Biol Chem 272, 25238-42.
(4) Y amamoto, K. et al. (1999) Mol Cell Biol 19, 8469-78.
-7
RP
M
I
Ju 82
rk 26
at
U2
66
HD
LM
KA -2
RP
L- AS54 29
9
0
HT
-2
9
**Anti-mouse secondary antibodies must be used to
detect this antibody.
kDa
200
RL
Background: Bcl-2 exerts a survival function in response
to a wide range of apoptotic stimuli through inhibition of
mitochondrial cytochrome c release (1). It has been implicated
in modulating mitochondrial calcium homeostasis and proton
flux (2). Several phosphorylation sites have been identified
within Bcl-2 including Thr56, Ser70, Thr74, and Ser87 (3). It
has been suggested that these phosphorylation sites may be
targets of the ASK1/MKK7/JNK1 pathway and that phosphorylation of Bcl-2 may be a marker for mitotic events (4,5). Mutation
of Bcl-2 at Thr56 or Ser87 inhibits its anti-apoptotic activity
during glucocorticoid-induced apoptosis of T lymphocytes (6).
Interleukin-3 and JNK-induced Bcl-2 phosphorylation at Ser70
may be required for its enhanced anti-apoptotic functions (7).
140
100
80
60
50
40
30
Bcl-2
20
10
60
50
Recommended Antibody Dilutions:
Western blotting
1:1000
Immunoprecipitation1:50
Immunohistochemistry (Paraffin)
1:400†
Unmasking buffer: EDTA
Antibody diluent: SignalStain® Antibody Diluent #8112
Detection reagent: SignalStain® Boost (HRP, Mouse) #8125
†Optimal IHC dilutions determined using SignalStain® Boost
IHC Detection Reagent.
Flow Cytometry
1:50
For product specific protocols and a complete listing
of recommended companion products please see the
product web page at www.cellsignal.com
β-Actin
40
Western blot analysis of extracts from various cell lines using
Bcl-2 (124) Mouse mAb (upper) and β-Actin (D6A8) Rabbit mAb
#8457 (lower). KARPAS cell Line source: Dr Abraham Karpas at the
University of Cambridge.
(5) L ing, Y.H. et al. (1998) J Biol Chem 273, 18984-91.
(6) H
uang, S.T. and Cidlowski, J.A. (2002) FASEB J 16, 825-32.
(7) D
eng, X. et al. (2001) J Biol Chem 276, 23681-8.
Events
Immunohistochemical analysis of paraffin-embedded human tonsil
using Bcl-2 (124) Mouse mAb.
Bcl-2
Immunohistochemical analysis of paraffin-embedded human lung
carcinoma using Bcl-2 (124) Mouse mAb.
Flow cytometric analysis of HT29 cells (blue), SHSY5Y cells (red)
and RL7 cells (green) using Bcl-2 Mouse mAb. Anti-mouse IgG
(H+L), F(ab')2 fragment (Alexa Fluor 488 Conjugate) #4412 was
used as a secondary antibody.
IMPORTANT: For western blots, incubate membrane
with diluted antibody in 5% w/v BSA, 1X TBS, 0.1%
Tween®20 at 4°C with gentle shaking, overnight.
Tween is a registered trademark of ICI Americas, Inc.
Thank you for your recent purchase. If you would like
to provide a review visit www.cellsignal.com/comments.
© 2015 Cell Signaling Technology, Inc.
SignalStain and Cell Signaling Technology are trademarks of Cell Signaling Technology, Inc.
Applications: W—Western IP—Immunoprecipitation IHC—Immunohistochemistry ChIP—Chromatin Immunoprecipitation IF—Immunofluorescence F—Flow cytometry E-P—ELISA-Peptide Species Cross-Reactivity: H—human M—mouse R—rat Hm—hamster
Mk—monkey Mi—mink C—chicken Dm—D. melanogaster X—Xenopus Z—zebrafish B—bovine Dg—dog Pg—pig Sc—S. cerevisiae Ce—C. elegans Hr—Horse All—all species expected Species enclosed in parentheses are predicted to react based on 100% homology.
Immunohistochemical analysis of paraffin-embedded RL-7 (positive, left) and HT-29 (negative, right) cell pellets using Bcl-2 (124)
Mouse mAb.
kDa
1
2
3
200
140
100
80
60
50
40
30
Bcl-2
20
10
–
+
–
Mouse (G3A1) mAb IgG1
Isotype Control
–
–
+
Bcl-2 (124) Mouse mAb
Immunoprecipitation of Bcl-2 from RL-7 cell extracts using
Mouse (G3A1) mAb IgG1 Isotype Control #5415 (lane 2) or
Bcl-2 (124) Mouse mAb (lane 3). Lane 1 is 10% input. Western
blot was performed using Bcl-2 (D55G8) Rabbit mAb (Human
Specific) #4223.
Orders: 877-616-CELL (2355) | [email protected] | Support: 877-678-TECH (8324) | www.cellsignal.com/support
© 2015 Cell Signaling Technology, Inc.
Immunohistochemical analysis of paraffin-embedded human breast
carcinoma using Bcl-2 (124) Mouse mAb.